Visualizing the Interface of Biotin and Fatty Acid Biosynthesis through SuFEx Probes

Aochiu Chen, Rebecca N. Re, Tony D. Davis, Kelley Tran, Yuta W. Moriuchi, Sitong Wu, James J. La Clair, Gordon V. Louie, Marianne E. Bowman, David J. Clarke, C. Logan Mackay, Dominic J. Campopiano, Joseph P. Noel, Michael D. Burkart*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

Site-specific covalent conjugation offers a powerful tool to identify and understand protein-protein interactions. In this study, we discover that sulfur fluoride exchange (SuFEx) warheads effectively crosslink the Escherichia coli acyl carrier protein (AcpP) with its partner BioF, a key pyridoxal 5′-phosphate (PLP)-dependent enzyme in the early steps of biotin biosynthesis by targeting a tyrosine residue proximal to the active site. We identify the site of crosslink by MS/MS analysis of the peptide originating from both partners. We further evaluate the BioF-AcpP interface through protein crystallography and mutational studies. Among the AcpP-interacting BioF surface residues, three critical arginine residues appear to be involved in AcpP recognition so that pimeloyl-AcpP can serve as the acyl donor for PLP-mediated catalysis. These findings validate an evolutionary gain-of-function for BioF, allowing the organism to build biotin directly from fatty acid biosynthesis through surface modifications selective for salt bridge formation with acidic AcpP residues.

Original languageEnglish
Pages (from-to)1388-1395
Number of pages8
JournalJournal of the American Chemical Society
Volume146
Issue number2
Early online date4 Jan 2024
DOIs
Publication statusE-pub ahead of print - 4 Jan 2024

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